CAJ | 학술논문

目的探讨并建立可供药物评价或生物学功能研究的表达人PSCA抗原的小鼠肿瘤模型.方法克隆人PSCA基因,构建pcDNA-PSCA质粒,稳定转染RM-1细胞,用RT-PCR和流式检测的方法筛选稳定表达人PSCA抗原的RM-PSCA细胞株;再将RM-PSCA细胞接种C57BL/6小鼠,观察其致瘤性,并寻找能够稳定致瘤的细胞数量;进而观测RM-PSCA所致肿瘤的生长情况及小鼠存活状况.结果筛选到了表达人PSCA抗原的RM-PSCA细胞,且1×10~5个肿瘤细胞能够保证10只实验小鼠全部成瘤;所致肿瘤生长迅速,接种后小鼠的平均存活时间为37 d.结论该研究成功的建立了稳定表达人PSCA抗原的小鼠肿瘤模型.
목적 탐토병건립가공약물평개혹생물학공능연구적표체인PSCA항원적소서종류모형.방법 극륭인PSCA기인,구건pcDNA-PSCA질립,은정전염RM-1세포,용RT-PCR화류식검측적방법 사선은정표체인PSCA항원적RM-PSCA세포주;재장RM-PSCA세포접충C57BL/6소서,관찰기치류성,병심조능구은정치류적세포수량;진이관측RM-PSCA소치종류적생장정황급소서존활상황.결과 사선도료표체인PSCA항원적RM-PSCA세포,차1×10~5개종류세포능구보증10지실험소서전부성류;소치종류생장신속,접충후소서적평균존활시간위37 d.결론 해연구성공적건립료은정표체인PSCA항원적소서종류모형.
Objective To establish a mouse model of prostate cancer expressing human PSCA for the development of new anti-tumor drugs or vaccines. Methods The total RNA of DU145 cells,a human prostate cancer cell line,was isolated by using TRIzol reagent according to the (RT-PCR),the first-strand cDNA was synthesized using the SuperScript First-Strand synthesis system. The human PSCA gene was amplified with the primers and cloned into the plasmid pcDNA3.1 to generate pcDNA-PSCA. DNA sequencing was used to confirm the constructs. The mouse prostate tumor cell line RM-1 cells,syngeneic to C57BL/6,were transfected with pcDNA-PSCA plasmids followed by selection using G418. RT-PCR analysis was performed to examine the validity of the constructs. Expression of PSCA on the cell surface was determined by staining with anti-PSCA antibody,and the anti-PSCA antibody was detected using an FITC-conjugated goat anti-rabbit IgG antibody,and analyzed by flow cytometry. 4-6-week-old male C57BL/6 mice purchased from the Laboratory Animals Center were inoculated with different amounts of RM-PSCA cells to search for suitable cell population which can form tumor in mouse,and the mice were monitored twice a week. The growth and the survival time of mice were measured,respectively. The tumor volume was measured by vernier caliper according to the formula:V=0.5a×b~2,where a and b are the long and short diameters of the tumor,respectively. Results The plasmid pcDNA-PSCA was successfully constructed and the PSCA was successfully expressed in RM-PSCA 7~# and RM-PSCA 28~# cells by RT-PCR and confirmed by flow cytometry. 1×10~5 RM-PSCA cells were sufficient to get tumor growth in 100% of inoculated mice. The tumor grew quickly and the volume of the tumor reached 12000 mm~3 within 34 days. All the mice died within 40 days and their mean survival time was 37 days. Conclusion A PSCA-expressing tumor model in mice has been successfully established. It can be used to evaluate the activities of drugs or vaccines.